Effect of electroacupuncture on autophagy of colonic cells in rats with ulcerative colitis
Objective To investigate the effect of electroacupuncture(EA)of"Changbing Decoction"on the level of autophagy in colonic cells in rats with ulcerative colitis(UC).Methods The UC model was established by using 5％dextran sulfate sodium(DSS)solution.Forty male SD rats were randomly divided into 5 groups:Rats in the normal control group(normal group),UC model group(model group),EA treatment group(EA group),UC group(RAPA group),and UC group(3-MA group)were treated with intraperitoneal injection of rapamycin(RAPA group),and UC group(3-MA group)were treated with intraperitone-al injection of 3-methyladenine(3-MA group).Ea was applied to"Zhongwan"(CV 12),bilateral"Tianshu"(ST 25)and"Shangjuxu"(ST 37)for 20 min,with disperse-dense wave,10 Hz/50 Hz in frequency and slight tremor in the limbs of rats.The body weight and colon length of rats were measured,and the disease activity index(DAI)score was calculated.The serum and colon tissue of rats in each group were collected,and the morphological changes of colon were observed by HE staining.The levels of tumor necrosis factor(TNF)-α,interleukin(IL)-6,IL-2 and IL-10 in serum were detected by enzyme-linked immunosorbent assay(ELISA).The ultrastructure of colonic mucosal epithelial cells was observed by ultramicroelectron micro-scope.Western blot was used to detect the relative levels of autophagy-related proteins(ATG 5,ATG 12,LC 3 and p 62).Quantitative real-time PCR(qPCR)was used to detect the mRNA levels of ATG 5,ATG 12,LC 3 and p 62.The UC model was established by using 5％dextran sulfate sodium(DSS)solution.Forty male SD rats were randomly divided into 5 groups:Rats in the normal control group(normal group),UC model group(model group),EA treatment group(EA group),UC group(RAPA group),and UC group(3-MA group)were treated with intraperitoneal injection of rapamycin(RAPA group),and UC group(3-MA group)were treated with intraperitoneal injection of 3-methyladenine(3-MA group).Ea was applied to"Zhongwan"(CV 12),bilateral"Tianshu"(ST 25)and"Shangjuxu"(ST 37)for 20 min,with disperse-dense wave,10 Hz/50 Hz in frequency and slight tremor in the limbs of rats.The body weight and colon length of rats were measured,and the disease activity index(DAI)score was calculated.The serum and colon tissue of rats in each group were collected,and the morphologi-cal changes of colon were observed by HE staining.The levels of tumor necrosis factor(TNF)-α,interleukin(IL)-6,IL-2 and IL-10 in serum were detected by enzyme-linked immunosorbent assay(ELISA).The ultrastructure of colonic mucosal epithelial cells was observed by ultramicroelectron microscope.Western blot was used to detect the relative levels of autophagy-related proteins(ATG 5,ATG 12,LC 3 and p 62).Quantitative real-time PCR(qPCR)was used to detect the mRNA levels of ATG 5,ATG 12,LC 3 and p 62.Results Compared with the normal group,the DAI scores of the model group and 3-MA group were increased,the colon length was shortened,the levels of IL-2 and IL-10 in serum were decreased,the levels of TNF-α and IL-6 were increased,and the relative protein expression levels and mRNA expression levels of ATG 5,ATG 12 and LC 3 in colon tissue were significantly decreased.The relative protein expression and mRNA expression of p62 were significantly in-creased(P＜0.05,P＜0.01).Compared with the model group,the DAI score was decreased,the colon length was increased,the levels of IL-2 and IL-10 in serum were increased,the levels of TNF-α and IL-6 were decreased,and the relative protein and mRNA levels of ATG 5,ATG 12 and LC 3 in colon tissue were significantly increased in the EA and RAPA groups.The rela-tive protein expression and mRNA expression of p 62 were significantly decreased(P＜0.05,P＜0.01).Compared with the EA group,the DAI score was increased,the colon length was shortened,the levels of IL-2 and IL-10 in serum were decreased,the levels of TNF-α and IL-6 in serum were increased,and the relative protein and mRNA levels of ATG 5,ATG 12 and LC 3 in colon tissue were significantly decreased in the 3-MA group.The relative protein expression of p62 and the relative mRNA expression of p62 were significantly increased(P＜0.05,P＜0.01).Conclusion EA of"Changbing Decoction"can promote the level of autophagy in colonic cells in rats with ulcerative colitis.
Ulcerative colitisElectroacupunctureAutophagy in cells
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